Cross-linking of mRNA to initiation factor eIF-3, 24 kDa cap binding protein and ribosomal proteins S1, S3/3a, S6 and S11 within the 48S pre-initiation complex

Nucleic Acids Res. 1984 Dec 11;12(23):8887-97. doi: 10.1093/nar/12.23.8887.

Abstract

Native small ribosomal subunits from rabbit reticulocytes contain all initiation factors necessary for the formation of the mRNA-containing 48S pre-initiation complex. The complex formed in the presence of Met-tRNAf and 125I-labelled globin mRNA was cross-linked with diepoxybutane, and the covalent mRNA-protein complexes were isolated under denaturating conditions. The proteins of the covalent complex were identified as the 110, 95 and 66/64 kDa subunits of eIF-3. In addition, the 24 kDa cap binding protein and the ribosomal proteins S1, S3/3a, S6 and S11 were found covalently linked to the mRNA. Ribosomal proteins S3/3a and S6 were also involved in the ribosomal mRNA-binding domain of reticulocyte polysomes.

MeSH terms

  • Animals
  • Carrier Proteins / metabolism*
  • Eukaryotic Initiation Factor-3
  • Globins / genetics
  • Peptide Chain Initiation, Translational
  • Peptide Initiation Factors / metabolism*
  • Polyribosomes / metabolism
  • Protein Binding
  • RNA Cap-Binding Proteins
  • RNA, Messenger / genetics
  • RNA, Messenger / isolation & purification
  • RNA, Messenger / metabolism*
  • Rabbits
  • Reticulocytes / metabolism
  • Ribosomal Proteins / isolation & purification
  • Ribosomal Proteins / metabolism*
  • Ribosomes / metabolism*
  • Structure-Activity Relationship

Substances

  • Carrier Proteins
  • Eukaryotic Initiation Factor-3
  • Peptide Initiation Factors
  • RNA Cap-Binding Proteins
  • RNA, Messenger
  • Ribosomal Proteins
  • Globins